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Methylation Profiling Using Methylated DNA Immunoprecipitation and Tiling Array Hybridization

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Book cover Germline Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 825))

Abstract

DNA methylation is an important epigenetic modification that regulates development and plays a role in the pathophysiology of many diseases. It is dynamically changed during germline development. Methylated DNA immunoprecipitation (MeDIP) is an efficient, cost-effective method for locus-specific and genome-wide analysis. Methylated DNA fragments are enriched by a 5-methylcytidine-recognizing antibody, therefore allowing the analysis of both CpG and non-CpG methylation. The enriched DNA fragments can be amplified and hybridized to tiling arrays covering CpG islands, promoters, or the entire genome. Comparison of different methylomes permits the discovery of differentially methylated regions that might be important in disease- or tissue-specific expression. Here, we describe an established MeDIP protocol and tiling array hybridization method for profiling methylation of testicular germ cells.

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References

  1. Weber M, Davies JJ, Wittig D, Oakeley EJ, Haase M, Lam WL, Schubeler D (2005) Chromosome-wide and promoter-specific analyses identify sites of differential DNA methylation in normal and transformed human cells, Nat Genet 37, 853–862.

    Article  PubMed  CAS  Google Scholar 

  2. Zhang X, Yazaki J, Sundaresan A, Cokus S, Chan SW, Chen H, Henderson IR, Shinn P, Pellegrini M, Jacobsen SE, Ecker JR (2006) Genome-wide high-resolution mapping and functional analysis of DNA methylation in arabidopsis, Cell 126, 1189–1201.

    Article  PubMed  CAS  Google Scholar 

  3. Weber M, Hellmann I, Stadler MB, Ramos L, Paabo S, Rebhan M, Schubeler D (2007) Distribution, silencing potential and evolutionary impact of promoter DNA methylation in the human genome, Nat Genet 39, 457–466.

    Article  PubMed  CAS  Google Scholar 

  4. Down TA, Rakyan VK, Turner DJ, Flicek P, Li H, Kulesha E, Graf S, Johnson N, Herrero J, Tomazou EM, Thorne NP, Backdahl L, Herberth M, Howe KL, Jackson DK, Miretti MM, Marioni JC, Birney E, Hubbard TJ, Durbin R, Tavare S, Beck S (2008) A Bayesian deconvolution strategy for immunoprecipitation-based DNA methylome analysis, Nat Biotechnol 26, 779–785.

    Article  PubMed  CAS  Google Scholar 

  5. Cokus SJ, Feng S, Zhang X, Chen Z, Merriman B, Haudenschild CD, Pradhan S, Nelson S F, Pellegrini M, Jacobsen SE (2008) Shotgun bisulphite sequencing of the Arabidopsis genome reveals DNA methylation patterning, Nature 452, 215–219.

    Article  PubMed  CAS  Google Scholar 

  6. Rauch T, Li H, Wu X, Pfeifer GP (2006) MIRA-assisted microarray analysis, a new technology for the determination of DNA methylation patterns, identifies frequent methylation of homeodomain-containing genes in lung cancer cells, Cancer Res 66, 7939–7947.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported in part by the Intramural Research Program of the National Institutes of Health (NIH), Eunice Kennedy Shriver National Institute of Child Health and Human Development, and in part by the Chinese University of Hong Kong.

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Correspondence to Hoi-Hung Cheung .

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© 2012 Springer Science+Business Media, LLC

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Cheung, HH., Lee, TL., Rennert, O.M., Chan, WY. (2012). Methylation Profiling Using Methylated DNA Immunoprecipitation and Tiling Array Hybridization. In: Chan, WY., Blomberg, L. (eds) Germline Development. Methods in Molecular Biology, vol 825. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-436-0_10

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  • DOI: https://doi.org/10.1007/978-1-61779-436-0_10

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-61779-435-3

  • Online ISBN: 978-1-61779-436-0

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